Passage and concentration-dependent effects of Indomethacin on tendon derived cells.

Mallick E, Scutt N, Scutt A, Rolf C - J Orthop Surg Res (2009)

Bottom Line:
As NSAIDs have been shown to delay healing in a number of mesenchymal tissues we have investigated the direct effects of indomethacin on the proliferation of tendon-derived cells.The results obtained were dependent on both the type of cells used and the method of measurement.These results suggest that firstly, care must be taken when deciding on methodology used to investigate tendon-derived cells as these cells have a quite different metabolism to other mesenchymal derive cells.

Background: Non-steroidal anti-inflammatory drugs (NSAID) are commonly used in the treatment of tendinopathies such as tendonitis and tendinosis. Despite this, little is known of their direct actions on tendon-derived cells. As NSAIDs have been shown to delay healing in a number of mesenchymal tissues we have investigated the direct effects of indomethacin on the proliferation of tendon-derived cells.

Results and discussion: The results obtained were dependent on both the type of cells used and the method of measurement. When measured using the Alamar blue assay, a common method for the measurement of cell proliferation and viability, no effect of indomethacin was seen regardless of cell source. It is likely that this lack of effect was due to a paucity of mitochondrial enzymes in tendon cells.However, when cell number was assessed using the methylene blue assay, which is a simple nuclear staining technique, an Indomethacin-induced inhibition of proliferation was seen in primary cells but not in secondary subcultures.

Conclusion: These results suggest that firstly, care must be taken when deciding on methodology used to investigate tendon-derived cells as these cells have a quite different metabolism to other mesenchymal derive cells. Secondly, Indomethacin can inhibit the proliferation of primary tendon derived cells and that secondary subculture selects for a population of cells that is unresponsive to this drug.

Figure 1: This graph shows the relationship between increasing Indomethacin concentration and cell number of primary tendon derived cells as measured by Alamar Blue.

Mentions:
Initial experiments studying the effects of indomethacin on tendon derived cell proliferation were carried out using the commonly used Alamar blue assay. However, although on visual examination an effect was evident, no effect was seen after analysis of the cell supernatants (Figure 1 & Figure 2).

Figure 1: This graph shows the relationship between increasing Indomethacin concentration and cell number of primary tendon derived cells as measured by Alamar Blue.

Mentions:
Initial experiments studying the effects of indomethacin on tendon derived cell proliferation were carried out using the commonly used Alamar blue assay. However, although on visual examination an effect was evident, no effect was seen after analysis of the cell supernatants (Figure 1 & Figure 2).

Bottom Line:
As NSAIDs have been shown to delay healing in a number of mesenchymal tissues we have investigated the direct effects of indomethacin on the proliferation of tendon-derived cells.The results obtained were dependent on both the type of cells used and the method of measurement.These results suggest that firstly, care must be taken when deciding on methodology used to investigate tendon-derived cells as these cells have a quite different metabolism to other mesenchymal derive cells.

Background: Non-steroidal anti-inflammatory drugs (NSAID) are commonly used in the treatment of tendinopathies such as tendonitis and tendinosis. Despite this, little is known of their direct actions on tendon-derived cells. As NSAIDs have been shown to delay healing in a number of mesenchymal tissues we have investigated the direct effects of indomethacin on the proliferation of tendon-derived cells.

Results and discussion: The results obtained were dependent on both the type of cells used and the method of measurement. When measured using the Alamar blue assay, a common method for the measurement of cell proliferation and viability, no effect of indomethacin was seen regardless of cell source. It is likely that this lack of effect was due to a paucity of mitochondrial enzymes in tendon cells.However, when cell number was assessed using the methylene blue assay, which is a simple nuclear staining technique, an Indomethacin-induced inhibition of proliferation was seen in primary cells but not in secondary subcultures.

Conclusion: These results suggest that firstly, care must be taken when deciding on methodology used to investigate tendon-derived cells as these cells have a quite different metabolism to other mesenchymal derive cells. Secondly, Indomethacin can inhibit the proliferation of primary tendon derived cells and that secondary subculture selects for a population of cells that is unresponsive to this drug.